DNAJB12 Overexpression Lysate

Western Blot: DNAJB12 Overexpression Lysate (Adult Normal) [NBL1-09934] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for DNAJB12.

• Reactivity: Hu
• Applications: WB

DNAJB12 Overexpression Lysate Summary

• Description:

  DNAJB12 Transient Overexpression Lysate

  
  Expression Host: HEK293T
  
  Plasmid: RC220679
  
  Accession#: NM_017626
  
  Protein Tag: C-MYC/DDK
  
  You will receive 1 vial of lysate (100ug), 1 vial of empty vector negative control (100ug), and 1 vial of 2xSDS sample buffer (250ul). Each vial of cell lysate contains 100ug of total protein (at 1 mg/ml). The 2xSDS Sample Buffer consists of 4% SDS, 125mM Tris-HCl pH6.8, 10% Glycerol, 0.002% Bromophenol blue, 100mM DTT.
• Gene: DNAJB12

Applications/Dilutions

• Dilutions:
  • Western Blot
• Application Notes: This product is intended for use as a positive control in Western Blot. Overexpression of the target protein was confirmed using an antibody to DDK (FLAG) epitope tag (NBP1-71705) present on the protein construct.
  
  Each vial of cell lysate contains 100ug of total protein which should be sufficient for 20-50 reactions. Depending on over-expression level, antibody affinity and detection system, some lysates can go as low as 0.1 ug per load. We recommend starting with 5ug of cell lysate. Add an equal amount of cell lysate and 2X SDS Sample buffer and boil the SDS samples for 10 minutes before loading.
• Theoretical MW: 41.7 kDa.
  Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.

Packaging, Storage & Formulations

• Storage: Store at -80C. Avoid freeze-thaw cycles.
• Buffer: RIPA buffer

Lysate Details for Array

• Type: Overexpression

Notes

HEK293T cells in 10-cm dishes were transiently transfected with a non-lipid polymer transfection reagent specially designed and manufactured for large volume DNA transfection. Transfected cells were cultured for 48hrs before collection. The cells were lysed in modified RIPA buffer (25mM Tris-HCl pH7.6, 150mM NaCl, 1% NP-40, 1mM EDTA, 1xProteinase inhibitor cocktail mix, 1mM PMSF and 1mM Na3VO4, and then centrifuged to clarify the lysate. Protein concentration was measured by BCA protein assay kit.

Alternate Names for DNAJB12 Overexpression Lysate

• DJ10FLJ20027
• DKFZp586B2023
• DnaJ (Hsp40) homolog, subfamily B, member 12
• dnaJ homolog subfamily B member 12
• FLJ0027

Background

DNAJB12 belongs to the evolutionarily conserved DNAJ/HSP40 family of proteins, which regulate molecular chaperone activity by stimulating ATPase activity. DNAJ proteins may have up to 3 distinct domains: a conserved 70-amino acid J domain, usually at the N terminus; a glycine/phenylalanine (G/F)-rich region; and a cysteine-rich domain containing 4 motifs resembling a zinc finger domain (Ohtsuka and Hata, 2000 [PubMed 11147971]).[supplied by OMIM]. Transcript Variant: This variant (2) differs in the 3' UTR compared to variant 1. Variants 1 and 2 both encode the same protein. CCDS Note: The coding region has been updated to start at an alternate in-frame start codon that is supported by transcript and conservation data. The upstream AUG translation initiation start site has a weak Kozak sequence and may be primate-specific while the downstream AUG has a stronger Kozak sequence and shows more conservation among mammals. Due to leaky scanning, it is possible that some ribosomes initiate translation at the downstream AUG while others initiate from the upstream AUG codon. The coding region was extended since there is no community standard or experimental evidence to show which is the preferred translation initiation site in humans.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Lysates are guaranteed for 6 months from date of receipt.

Bioinformatics

• Gene Symbol: DNAJB12